Experimental and Theoretical Studies of Green Synthesized Cu2O Nanoparticles Using Datura Metel L.

In biomedical applications, Cu2O nanoparticles are of great interest. The bioengineered route is eco-friendly for the synthesis of nanoparticles. Therefore, in the present study, there is an attempt to synthesis Cu2O nanoparticles using Datura metel L. The synthesized nanoparticles were characterized by UV-Vis, XRD, and FT-IR. UV-Vis results suggest the presence of hyoscyamine, atropine in Datura metel L, and also, nanoparticles formation has been confirmed by the presence of absorption peak at 790 nm. The average crystallite size (19.56 nm) was obtained by XRD. FT-IR was also used to confirm the different functional groups. Fourier Power Spectrum was also employed to examine the synthesized nanomaterials spectrum data to emphasize the peak of the prominent frequencies. Density functional theory (DFT) was also utilized to assess the energy of the substance over time, which appears to indicate a stable molecule. Furthermore, calculated energies, thermodynamic properties (such as enthalpies, entropies, and Gibbs-free energies), modeled structures of complexes, crystals, and clusters, and predicted yields, rates, and regio- and stereospecificity of reactions were all in good agreement with experimental results. Overall, the results show that the successful production of Cu2O nanoparticles with Datura metel L. corresponds to theoretical research.

New withanolides with anti-inflammatory activity from the leaves of Datura metel L.

Twenty three undescribed withanolides, daturmetelides A-W (1-23), were isolated from 70% EtOH extract of the leaves of Datura metel L. The structural characterizations and relative configurations of 1-23 were elucidated by extensive spectroscopic analysis as well as by comparison with literature values. The absolute configurations of 1 and 3 were determined by X-ray crystallography. Bioassay results showed that 1 and 7 exhibited moderate inhibitory effects against NO production in lipopolysaccharide-stimulated RAW 264.7 cells (IC50 values of 13.74 µM and 13.92 µM, respectively). In addition, 1 and 7 showed significant anti-inflammatory activities against the production of TNF-α, IL-1ß, IL-6 and COX-2. Western blot analysis was further performed to reveal the mechanism of anti-inflammatory action via inhibition of the NF-κB activation.

Spleen and thymus metabolomics strategy to explore the immunoregulatory mechanism of total withanolides from the leaves of Datura metel L. on imiquimod-induced psoriatic skin dermatitis in mice.

Our previous work demonstrated that total withanolides of Datura metel L. leaves (TWD) exhibited excellent therapeutic effects on psoriasis. However, current knowledge of its mechanisms is incomplete. In this study, integrated spleen and thymus untargeted metabolomics were used to analyze the changes in endogenous metabolites underlying the immunosuppressive activity of TWD on psoriasis animal models induced by imiquimod. The results suggested that TWD treatment markedly attenuated imiquimod-induced psoriasis and showed significant immunosuppressive activity as evidenced by decreased elevation index of spleen and thymus. Meanwhile, TWD significantly reversed the elevation of immunoregulatory factors, including IL-10, IL-17, IL-22 and IL-23. Multivariate trajectory analysis revealed that TWD treatment could restore the psoriasis-disturbed spleen and thymus metabolite profiles towards the normal metabolic status. A total of 25 and 27 metabolites associated with the immunomodulatory effects for which levels changed markedly upon treatment have been identified in spleen and thymus, respectively. These differential metabolites were mainly involved in amino acid metabolism, nucleotide metabolism, fatty acid metabolism and lipid metabolism. Our investigation provided a holistic view of TWD for intervention in psoriasis through immunoregulation and provided further scientific information in vivo about a clinical value of TWD for psoriasis.

UPLC-MS/MS Identification and Quantification of Withanolides from Six Parts of the Medicinal Plant Datura Metel L.

Withanolides from six parts (flower, leaf, stem, root, seed, and peel) of Datura metel L. (D metel L.) obtained from ten production areas in China were identified and quantified by UPLC-MS/MS. A total of 85 withanolides were characterized for the first time using the UPLC-Q-TOF-MS/MS system. Additionally, a simultaneous, rapid and accurate measurement method was developed for the determination of 22 bioactive withanolides from ten production areas with the UPLC-Q-TRAP-MS/MS system. The results show the total withanolide content is highest in the leaves (155640.0 ng/g) and lowest in the roots (14839.8 ng/g). Compared with other production areas, the total content of plants from Dujiangyan was the highest at 82013.9 ng/g (value range of ten areas: 82013.9-42278.5 ng/g). The results also show significant differences in the distribution of withanolides in the different plant parts, as well as across different production areas. This is a breakthrough report providing a simultaneous qualitative and quantitative analysis of 22 withanolides in D. metel L. It could be the basis for the more rational use of various parts of D. metel L., and the expansion of medicinal resources. This work also lays a solid foundation for research on the quality control of D. metel L.

Daturanolide A-C, Three New Withanolides from Datura metel L. and Their Cytotoxic Activities.

Three new withanolides (1-3), named as daturanolide A-C, along with six known withanolides (4-9) were isolated from the flowers of Datura metel L. Their structures with absolute configurations were elucidated by a series of spectroscopic methods, electronic circular dichroism (ECD) analyses, and X-ray crystallography. All the isolates were evaluated for cytotoxicity against five human cancer cell lines (HCT116, U87-MG, NCI-H460, BGC823, and HepG2), and 6 exhibited marked cytotoxicity.

Withanolides, Extracted from Datura Metel L. Inhibit Keratinocyte Proliferation and Imiquimod-Induced Psoriasis-Like Dermatitis via the STAT3/P38/ERK1/2 Pathway.

Psoriasis is an immune-mediated inflammatory dermatosis characterized by epidermal hyperplasia and excessive infiltration of inflammatory cells. Withanolides, extracted from Datura metel L.; are the main effective components for the treatment of psoriasis. However, the precise mechanisms of action of withanolides for the treatment of psoriasis remain unclear. We found that treatment with withanolides alleviated imiquimod (IMQ)-induced epidermal hyperplasia and inflammatory cell infiltration in the effective skin of model mice. In addition, we also found that withanolides suppressed the activation of STAT3, ERK1/2 and P38 signaling pathways in IMQ-stimulated HaCat cells. These results suggest that withanolides possess an anti-inflammatory effect and have significant therapeutic potential for the prevention and treatment of psoriasis.

Datura Metel L. Ameliorates Imiquimod-Induced Psoriasis-Like Dermatitis and Inhibits Inflammatory Cytokines Production through TLR7/8-MyD88-NF-κB-NLRP3 Inflammasome Pathway.

BACKGROUND: Psoriasis is a chronic, immune-mediated inflammatory skin disease, and the inflammatory response plays an important role in its development and progression. Datura metel L. is a traditional Chinese medicine that exhibited a significant therapeutic effect on psoriasis in our previous study due to its remarkable anti-inflammatory effect. Meanwhile, the mechanism underlying its effects on psoriasis is still unclear. METHODS: An imiquimod-induced psoriasis-like dermatitis mouse model was constructed to evaluate the protective effect of the effective part of Datura metel L. (EPD), which was verified by evaluations of the Psoriasis Area and Severity Index (PASI) score. Hematoxylin and eosin (H&E) staining, immunohistochemical examination, enzyme-linked immunosorbent assay (ELISA), and Western blot were used to measure the inflammatory cytokines and the protein expression associated with the Toll-like receptor 7- myeloid differentiation primary response gene 88-nuclear Factor-κB-nucleotide-binding oligomerization domain (Nod)-like receptor family pyrin domain-containing 3 (TLR7/8-MyD88-NF-κB-NLRP3) inflammasome pathway. RESULTS: EPD significantly decreased the PASI, reduced epidermal thickness, and decreased the proliferation and differentiation of epidermal cells in psoriasis-like dermatitis C57BL/6 mice induced by imiquimod (IMQ). Furthermore, EPD reduced the infiltration of CD3+ cells to psoriatic lesions, as well as ameliorated the elevations of intercellular adhesion molecule 1 (ICAM-1) and inhibited the production of imiquimod-induced inflammatory cytokines, including IL-1ß, IL-2, IL-6, IL-10, IL-12, IL-17, IL-22, IL-23, tumor necrosis factor-α (TNF-α), monocyte chemotactic protein 1 (MCP-1), and interferon-γ (IFN-γ). Besides, EPD decreased the imiquimod-induced expression levels of TLR7, TLR8, TRAF6, MyD88, p-IKKα, p-IKBα, p-NF-κB, NLRP3, apoptosis-associated speck-like protein contained a caspase recruitment domain (ASC), cysteinyl aspartate specific proteinase 1 (caspase-1), and IL-1ß. CONCLUSION: This study demonstrated that EPD exhibited a protective effect on an imiquimod-induced psoriasis mice model by inhibiting the inflammatory response, which might be ascribed to the inhibition of the TLR7/8-MyD88-NF-κb-NLRP3 inflammasome pathway.

Datura metel L. leaf extract mediated sodium alginate polymer membrane for supercapacitor and food packaging applications.

Datura metel L. leaf extract mediated sodium alginate polymer membrane was successfully made using the solution casting technique. Electric, electrochemical, physicochemical and antimicrobial analyses of the prepared film were investigated. Functional groups of polysaccharides are identified in FTIR analysis and crystallinity/amorphous of the prepared samples was studied using XRD analysis. The prepared polymer membrane (DmMSA2) exhibits the ionic conductivity of 2.18 × 10-4 Scm-1, maximum specific capacitance of 131 F/g at a current density of 0.2 A/g and also exhibits a significant effect of antimicrobial activity against human pathogens. Hence, Datura metel L. leaf extract mediated polymer membranes are promising candidates for solid-electrolyte in supercapacitor devices and antimicrobial agents in food packaging applications.

Seven new glycosides from the leaves of Datura metel L.

Seven new glycosides (1-7) and seventeen known analogues (8-24) were isolated from the leaves of Datura metel L. The structures of these compounds were all elucidated by detailed spectroscopic analyses and comparison with literature values. All isolates were evaluated for cytotoxicity against Hela, MGC-803, Ishikawa cell lines and compounds 6, 7, 14-17 and 24 exhibited different degrees of antiproliferative effects.

New steroids from the pericarps of Datura metel L.

Three new steroids (1-3) and 13 reported analogs (4-16) were extracted from Datura metel L. pericarps. Structure analysis of these extracted compounds was performed by 1 D-NMR and 2 D-NMR spectroscopy, and their spectra were compared with those of similar compounds previously described in the literature. The extracted steroids (1-3) and known compounds (4-16) were evaluated for anti-inflammatory activity against LPS-activated RAW 264.7 cells. Compounds 5, 7, 9, 12 and 15 showed potential anti-inflammatory activity with IC50 less than 35 µM, while compounds 3 and 11 showed weak anti-inflammatory activity with IC50 less than 100 µM.

Total withanolides ameliorates imiquimod-induced psoriasis-like skin inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Datura metel L. has been used as an anesthetic in clinic for more than 1800 years in China, and the main efficacy of D. metel L. flower is relieving asthma and cough, relieving spasm and relieving pain. From 1978 to 1980, Datura metel L. was used as an anesthetic agent and occasionally cured psoriasis patients during anesthesia clinically, and our group confirmed that the effective portion is total withanolides (YWS). Moreover, the new drug "Datura metel L. capsule" composed of YWS has since been approved and used for the treatment of more than 3,000 psoriasis patients, with efficacy and cure rates greater than 90% and 65%. However, the immunological mechanism has not been elucidated. AIM OF THE STUDY: Nowadays, although total withanolides from Datura metel L. have a better clinical efficacy in the treatment of psoriasis, there is a lack of overall understanding of the mechanism of their treatment, especially about some immune cells and proteins closely related to psoriasis and their relationship in executive function and biological significance. This study focused on investigating the mechanism of psoriasis treatment by YWS and determined the biochemical processes in the treatment of psoriasis based on Treg/Th17 axis cell-mediated bidirectional immunoregulatory functions, which provides an important scientific basis for understanding the mechanism underlying the treatment of psoriasis by YWS. MATERIALS AND METHODS: The effects of YWS on the lesion pathology of IMQ-induced psoriasis mice and the underlying molecular mechanism were assessed directly using HE staining, the PASI score and the animal body mass. We also investigated the effects of YWS on the Treg/Th17 axis and their critical functions in psoriasis pathogenesis via molecular biological methods. Finally, we performed differential proteomics analysis on skin in IMQ-induced psoriasis mice to clarify the effect of YWS by incorporates mass spectrometry-bioinformatics and annotated the functions and pathways associated with the differential proteins through GO enrichment, KEGG pathway analysis and PPI networks analysis, respectively. RESULTS: YWS regulated the imbalance of the Treg/Th17 axis. And proteomic analysis showed that YWS up-regulated 46 and down-regulated 37 proteins. According to the bioinformatics analysis, the improvement of Treg/Th17 imbalance may be the key immunological mechanism of YWS in the treatment of psoriasis by up-regulating the butyrate metabolism pathway, down-regulating leukocyte migration, inhibiting the phagocytic function of natural killer cells, suppressing osteoclast differentiation and interfering with chemokine activity, and the critical proteins involved are Lyn, HMGCS2, ABAT, ITGß2, PRKCß, MMP9, NCF1, JUNß, and Hck. CONCLUSION: This research clarified that the improvement of the imbalance of the Treg/Th17 axis may be the key immunological mechanism of YWS in the treatment of psoriasis through metabolic pathways and influencing key proteins. The results not only expand the therapeutic targets and approaches for the treatment of psoriasis, which is a challenging and complex disease, but also deepens the understanding of the mechanism of YWS in the treatment of psoriasis and other important conditions to open up a new way of thinking for research on YWS in the treatment of psoriasis.

A new sesquiterpenoid with cytotoxic and anti-inflammatory activity from the leaves of Datura metel L.

A new sesquiterpenoid glycoside, consisting of one vetispirane-type, dmetelisproside A (1), together with two known megastigmane sesquiterpenoids, citroside A (2) and staphylionoside D (3) were isolated from the leaves of Datura metel L. The structures were determined by analysis of their MS and NMR data as well as by comparison with literature values. All isolates were evaluated for cytotoxicity toward SGC-7901, Hepg2, HeLa, MCF-7, and MDA-MB-231 cancer cell lines, and anti-inflammatory activity against nitric oxide (NO) in RAW 264.7 cells in vitro. Compounds 1 and 2 exhibited pronounced cytotoxicity against SGC-7901 and HeLa cells with IC50 values in the range of 21.43 to 29.51 µm. All three compounds displayed potential effects against NO production with IC50 values of 31.10, 34.25, and 44.31 µm, respectively. Thus, these isolated compounds in the leaves of Datura metel L were subjected to bioactivity substances for future studies on medicinal application.

UPLC-orbitrap-MS-based metabolic profiling of HaCaT cells exposed to withanolides extracted from Datura metel.L: Insights from an untargeted metabolomics.

In recent decades, more and more attention to the withanolides extracted from Datura metel.L has been paid due to their anti-psoriatic effects. Withanolides have also been reported to exhibit anti-inflammatory and anti-proliferative properties. Thus, withanolides have been considered as a promising candidate of anti-psoriatic drug. The aim of this study was to investigated the metabolic network of HaCaT cells after exposure to withanolides to identify anti-psoriatic mechanism induced by withanolides on skin cells. In this experiment, our results demonstrated that exposure to withanolides at concentrations beyond 50 µg/mL inhibited cell proliferation and induced cell apoptosis in a dose-dependent manner. In addition, withanolides-induced reactive oxygen species (ROS) generation and mitochondrial depolarization in HaCaT cells. In this research, ultra-high performance liquid chromatography coupled with orbitrap mass spectrometry (UPLC-orbitrap-MS) method was applied to profile metabolite changes in HaCaT cells exposed to withanolides. In total, significant variations in 38 differential metabolites were identified between withanolides exposure and untreated groups. The exposure of HaCaT cells to withanolides at the dose of 200 µg/mL for 24 h was revealed by the disturbance of energy metabolism, amino acid metabolism, lipid metabolism and nucleic acid metabolism. UPLC-orbitrap-MS-based cell metabolomics provided a comprehensive method for the identification of withanolides' anti-psoriasis mechanisms in vitro. And above metabolic disorders also reflected potential therapeutic targets for treating psoriasis.

Seven undescribed steroids from the leaves of Datura metel L.

Extraction of Datura metel L. leaves with ethanol as a solvent gave a group of steroids, including two unique 1,10-seco-withanolides (1, 4), an unusual nitrogen-containing withanolides (2), one undescribed saponin (3), two withanolides with a carbohydrate (5, 6), and one C21 steroid (7). These compounds' structures were identified based on HR-ESI-MS and 1H, 13C NMR data analyses, also compared with data from the document. Some compounds showed moderate inhibition on NO production in lipopolysaccharide-stimulated RAW 264.7 cells.

Integrated serum metabolomics and network pharmacology approach to reveal the potential mechanisms of withanolides from the leaves of Datura metel L. on psoriasis.

Psoriasis is a chronic, immune-mediated inflammatory skin disease and highly depends on inflammation and angiogenesis as well as other pathways. Our previous study showed that the withanolides from the leaves of Datura metel L. exhibited significant therapeutically effect on psoriasis, but the mechanisms concerning this effect have not been systematically studied. The purpose of this paper was to investigate the possible mechanism of withanolides for treating psoriasis using an integrated metabolomics and network pharmacology strategy. Untargeted metabolomics profiling of serum with UHPLC/Orbitrap MS and a multivariate data method were performed to discover the potential biomarkers and metabolic pathways. Afterward, the compound-target-pathway network of withanolides for psoriasis was constructed by virtue of network pharmacology. Finally, the crucial pathways were selected by integrating the results of metabolomics and network pharmacology, and then validated by ELISA and western blot analysis. The results showed that withanolides could exert excellent effects on psoriasis through regulating two types of pathways, angiogenesis and inflammation, including sphingolipids metabolism and HIF-1α/VEGF pathway, reflected by inhibiting the production of inflammatory cytokines (IL-1ß, IL-6, IL-8, IFN-γ, TNF-α, HIF-1α and VEGF), as well as reducing the protein expressions of HIF-1α and VEGF. Our study successfully explained the polypharmcological mechanisms underlying the efficiency of withanolides from the D. metel L. leaves on treating psoriasis. Meanwhile, it was also valuable for performing a systematical investigation of herb medicines, as well as for efficiently predicting the therapeutic mechanisms of traditional Chinese medicine.

Immunosuppressive withanolides from the flower of Datura metel L.

Thirteen new withanolide aglycones, baimantuoluolines L-X (1-13) and one new withanolide glycoside, baimantuoluoside J (14) were isolated from Datura metel L. flowers. The structures of the new compounds were elucidated by the detailed analysis of 1D and 2D NMR techniques and mass spectrometry, together with the closely related literatures. Meanwhile, all isolated compounds were evaluated for their immunosuppressive activities against mice splenocyte proliferation and antiproliferative activities against human gastric adenocarcinoma cells (SGC-7901), human hepatoma (HepG2), and human breast cancer (MCF-7) in vitro. It was found that compounds 1-14 showed obvious immunosupressive effects and some of them have moderated antiproliferative activities.

Anti-inflammatory sesquiterpenoids from the leaves of Datura metel L.

Nine new (1-9) and three known (10-12) sesquiterpenoids were isolated from the ethanol-water (7:3, v/v) extract of the Datura metel L. leaves. The structures of 1-9 were elucidated by detailed spectroscopic analyses, including 1D and 2D NMR, HR-ESI-MS. All isolates (1-12) were evaluated for anti-inflammatory activity against the production of nitrogen oxide in lipopolysaccharide-induced RAW264.7 cells and compound 5 possessed the best inhibitory effect among them, with the IC50 value reaching 9.33-11.67 µM, which was lower than positive control, L-NMMA, with IC50 range from 13.64 to 17.02 µM.

Daturmetesides A-E, five new ergostane-type C28 sterols from the leaves of Datura metel L.

Five undescribed ergostane-type C28 sterols, daturmetesides A-E (1-5), were isolated from the leaves of Datura metel L. The chemical structures of these new compounds were characterized through extensive spectroscopic analysis and comparison with literatures. Among them, the absolute structures of daturmetesides A and C were unambiguously determined by X-ray crystallography. The anti-inflammatory effect of daturmetesides A-E was all tested by measuring nitric oxide production in lipopolysaccharide-activated RAW264.7 cells. Daturmetesides A, C and D moderatelylowered the NO production with IC50 values ranging from 17.05 ± 0.35 to 24.88 ± 0.93 µM.

Withanolides from the leaves of Datura metel L.

Twelve previously undescribed withanolides, including three uncommon chemotypes of aglycones, daturafolisides (J-L), and six common structures, daturafolisides (M-U), as well as one known withanolide were isolated from the leaves of Datura metel L. Their structures were elucidated by interpretation of spectroscopic data (1D, 2D NMR and CD), HRESIMS experiments and comparison with published data for similar compounds. These isolated compounds were evaluated for their inhibitory activities on nitric oxide production. The IC50 values of daturafoliside L, daturafoliside M, and daturafoliside T were between 9.37 and 18.64 µM, while daturafoliside K, daturafoliside R, and daturametelin A had IC50 values of 22.83-33.36 µM. Herein, three proposed biosynthetic pathways for uncommon structures daturafolisides J-L were discussed. These constituents found in the leaves of Datura metel L. may be a good source of bioactive substances, making contributions to the sustainable utilization of this plant resources.

Chemical composition and cytotoxicity of the essential oil from different parts of Datura metel L.

The essential oil from different parts of Datura metel L. were extracted using hydrodistillation and GC-MS was used to analyse the essential oil. The main components of flowers were ketone (23.61%) and ethyl palmitate (15.84%). The main components of leaves were ketone (18.84%) and phytol (18.71%). Ketone (39.45%) and phytol (31.32%) were the major components of petioles. Palmitic acid (30.60%) and ethyl linoleate (21.56%) were the major components of seeds. The major ingredient of roots was palmitic acid (52.61%). The main ingredients of the stems were palmitic acid (38.38%) and ethyl linoleate (17.38%). All the different parts of essential oil were screened for cytotoxicity. The roots and stems showed the inhibitory effects against HepG-2 with IC50 levels of 613.88 and 341.12 mg/L. The leaves and roots showed the inhibitory effects against HeLa with IC50 levels of 267.76 and 348.35 mg/L. All the six parts have inhibitory effects against SGC-7901 cell lines.